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Mouse CD40L(Cluster of Differentiation 40 Ligand) ELISA Kit
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Product No

FY-EM5394

Sample Type

Serum, Plasma, Tissue homogenate and Other biological samples

Assay Time

3.5H

Price($)

280 400

Size

48T 96T

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DataSheet DataSheet MSDS MSDS
Product Details
Product Name in English:
Mouse CD40L(Cluster of Differentiation 40 Ligand) ELISA Kit
Species:
Mouse
Alternative Name:
TNFSF5, CD4LG, CD154, HIGM1, IGM, IMD3, T-BAM, TRAP, Gp39
Uniport ID:
P27548
Assay Type:
Sandwich
Standard:
6000pg/mL
Sensitivity:
56.25 pg/mL
Detection Range:
93.75-6000pg/mL
Assay Time:
3.5H
Sample Type:
Serum, Plasma, Tissue homogenate and Other biological samples
Research:
Cardiovascular biology
Test Principle:
The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to CD40L. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to CD40L. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain CD40L, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of CD40L in the samples is then determined by comparing the OD of the samples to the standard curve.
Technical Data
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
Concentration OD1 OD2 Average OD Corrected
6000 2.761 2.782 2.772 2.731
3000 1.699 1.712 1.706 1.669
1500 0.962 0.966 0.964 0.932
750 0.433 0.434 0.434 0.403
375 0.215 0.216 0.216 0.185
187.5 0.108 0.108 0.108 0.078
93.75 0.0629 0.063 0.063 0.0329
0 0.03 0.03 0.03 0
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level were tested on 3 different plates, 20 replicates in each plate, respectively.
Item Intra-assay Precision Intra-assay Precision
Sample 3 3
Replicate 9 18
CV(%) 5 8
Rate of recovery
The recovery of spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
Sample Type Range(%) Average Recovery(%)
Serum (n=8) 90-103 96
EDTA plasma (n=8) 81-96 88
Cell culture media (n=8) 80-94 86
Linear
Samples were spiked with high concentrations of target proteins and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
1:2 Range (%) 86-96 87-96 87-96
Average (%) 89 95 88
1:4 Range (%) 93-104 87-96 85-97
Average (%) 93 95 87
1:8 Range (%) 89-97 85-97 87-96
Average (%) 91 89 96
1:16 Range (%) 85-94 85-94 85-96
Average (%) 90 90 86
Assay Procedures
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